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9776
Myosin Light Chain 2 Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Myosin Light Chain 2 Antibody Sampler Kit #9776

Citations (5)

Simple Western™ analysis of lysates (0.1 mg/mL) from C2C12 cells using Phospho-Myosin Light Chain 2 (Thr18/Ser19) Antibody #3674. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 2 – 40 kDa separation module.

Western blot analysis of extracts from HeLa cells, vehicle-treated (-) or treated with the myosin light chain kinase inhibitor ML-7 (50μM, 15 min; +) using Phospho-Myosin Light Chain 2 (Ser19) Antibody (upper) or Myosin Light Chain 2 (D18E2) Rabbit mAb #8505 (lower).
Western blot analysis of extracts from HeLa cells, vehicle-treated (-) or treated with the myosin light chain kinase inhibitor ML-7 (50μM, 15 min; +) using Phospho-Myosin Light Chain 2 (Thr18/Ser19) Antibody (upper) or Myosin Light Chain 2 (D18E2) Rabbit mAb #8505 (lower).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from various cell lines using Myosin Light Chain 2 (D18E2) Rabbit mAb.
Confocal immunofluorescent images of HeLa cells, untreated (left) or phosphatase-treated (right), labeled with Phospho-Myosin Light Chain 2 (Ser19) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
To Purchase # 9776
Cat. # Size Qty. Price
9776T
1 Kit  (3 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-Myosin Light Chain 2 (Ser19) Antibody 3671 20 µl
  • WB
  • IF
H M R Dm 18 Rabbit 
Phospho-Myosin Light Chain 2 (Thr18/Ser19) Antibody 3674 20 µl
  • WB
H M 18 Rabbit 
Myosin Light Chain 2 (D18E2) Rabbit mAb 8505 20 µl
  • WB
H M R 18 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Kit Usage Information

Protocols

Product Description

The Myosin Light Chain 2 Antibody Sampler Kit provides an economical means to detect total, phosphorylated, and dual-phosphorylated myosin light chain 2. The kit contains enough primary and secondary antibody to perform two western blot experiments.

Specificity / Sensitivity

The antibodies in the Myosin Light Chain 2 Antibody Sampler Kit detect endogenous levels of total myosin light chain 2 (smooth muscle), myosin light chain 2 when phosphorylated at Ser19 or when dually phosphorylated at Thr18 and Ser19, respectively. The antibodies do not cross-react with the cardiac isoform of Myosin Light Chain 2.

Source / Purification

The rabbit monoclonal antibody is produced by immunizing rabbits with a synthetic peptide corresponding to residues near the carboxy terminus of human myosin light chain 2 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser19 of human myosin light chain 2, or to residues surrounding Thr18/Ser19 of human myosin light chain 2, respectively. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Myosin is composed of six polypeptide chains: two identical heavy chains and two pairs of light chains. Myosin light chain 2 (MLC2), also known as myosin regulatory light chain (MRLC), RLC, or LC20, has many isoforms depending on its distribution. In smooth muscle, MLC2 is phosphorylated at Thr18 and Ser19 by myosin light chain kinase (MLCK) in a Ca2+/calmodulin-dependent manner (1). This phosphorylation is correlated with myosin ATPase activity and smooth muscle contraction (2). ROCK also phosphorylates Ser19 of smooth muscle MLC2, which regulates the assembly of stress fibers (3). Phosphorylation of smooth muscle MLC2 at Ser1/Ser2 and Ser9 by PKC and cdc2 has been reported to inhibit myosin ATPase activity (4,5). Phosphorylation by cdc2 controls the timing of cytokinesis (5). Transgenic mice lacking phosphorylation sites on the cardiac muscle isoform show morphological and functional abnormalities (6).

Pathways

Explore pathways related to this product.

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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